Isolation of photoreceptors in the cultured full-thickness fetal rat retina.
OBJECTIVE
To create a retina consisting mainly of photoreceptors for future use as donor tissue in retinal transplantation.


METHODS
Fetal full-thickness neuroretinas from Sprague-Dawley embryonic day (E) 17 or E20 rats were placed in culture for 7 or 14 days. Explants and age-matched control retinas were examined by light microscopy and with a panel of immunohistochemical markers labeling all seven of the major retinal cell types.


RESULTS
E17 and E20 control retinas displayed vimentin-labeled Müller cells, NF160-labeled ganglion cells, and synaptic vesicles labeled with synaptophysin. The remaining cell types were found in control specimens of postnatal age 2 days and older. After 7 or 14 days in culture, all explants were significantly thinner than their aged-matched controls and displayed multiple rows of cells organized in a single layer. Within this layer, they contained rhodopsin-labeled rod photoreceptors, presynaptic vesicles, and vertically arranged Müller cells. Transducin-labeled cone photoreceptors were found in all but the youngest explants. Scattered PKC-labeled rod bipolar cells and calbindin-labeled horizontal cells were found in the inner part of most explants, whereas beta-III-tubulin-labeled ganglion cells and parvalbumin-labeled amacrine cells were seen only sporadically. No NF160-labeled ganglion cells were found.


CONCLUSIONS
Fetal full-thickness rat retina in vitro develops into a retina consisting of predominantly synapse-containing cone and rod photoreceptors embedded in a scaffold of well-organized Müller cells. These explant retina characteristics are well adapted for use as donor tissue in future retinal transplantation experiments.
Ghosh
Fredrik
F
Department of Ophthalmology, Lund University Hospital, Lund, Sweden. fredrik.ghosh@med.lu.se
Arnér
Karin
K
Engelsberg
Karl
K
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology; D002469:Cell Separation; D002470:Cell Survival; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D008297:Male; D009457:Neuroglia / Q000166:cytology / Q000378:metabolism; D009924:Organ Culture Techniques; D020419:Photoreceptor Cells, Vertebrate / Q000166:cytology* / Q000378:metabolism; D011247:Pregnancy; D051381:Rats; D017207:Rats, Sprague-Dawley; D012160:Retina / Q000196:embryology* / Q000378:metabolism; D051245:Retinal Bipolar Cells / Q000166:cytology / Q000378:metabolism; D012165:Retinal Ganglion Cells / Q000166:cytology / Q000378:metabolism; D013572:Synaptic Vesicles / Q000378:metabolism
D015415:Biomarkers